MEASLES VIRUS STUDIES IN A TRANSGENIC MODEL

转基因模型中的麻疹病毒研究

• 批准号: 2072368
• 负责人: MICHAEL B OLDSTONE
• 金额: $ 23.48万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-09-30 至 1997-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2072368
• 关键词: CD antigens; active immunization; antiviral antibody; biological models; cell mediated cytotoxicity; cell mediated lymphocytolysis test; cellular immunity; cytotoxic T lymphocyte; epitope mapping; flow cytometry; gene expression; genetically modified animals; histocompatibility antigens; humoral immunity; immunodeficiency; laboratory mouse; major histocompatibility complex; measles; measles vaccine; measles virus; model design /development; tissue /cell culture; virulence; virus infection mechanism; virus receptors

Measles virus causes significant morbidity and mortality in the human population. Despite a successful attenuated vaccine, measles virus still kills over one million children a year. The problem being that the vaccine is inefficient in children infected prior to their ninth month of age. This is believed due to the presence of maternal antibody and/or virus induced immunosuppression or other late effects following vaccination. Further, measles causes a hyperacute allergic disease and a chronic persistent (subacute sclerosing panencephalitis) neuronal disorder. Yet our understanding and knowledge of the molecular features of measles virus, the immunologic and immunopathologic responses to both infection and vaccination and how the virus persists in neurons are poorly understood. In order to understand the pathogenesis of measles virus infection and to identify the immune protective response, including B and T cell epitopes, we will develop and exploit a novel small animal model. We and our colleagues have identified the putative receptor for measles virus, the membrane cofactor protein (MCP, CD46). Measles virus is permissive for human and simian but not murine cells. However, murine MC57 and 3T3 cells barely permissive to measles virus become permissive and produce progeny virus when stably expressing any of the four CD46 isoforms, although isoform BC1 and BC2 are associated with a higher degree of syncytial formation. We propose expressing the cDNA of CD46 in mice under its own, a beta actin (universal) and cell-specific (neuron specific enolase, RIP) promoters. This will allow us to generate a transgenic model of measles virus in a small, inexpensive, genetically manipulable host, the mouse, whose immunologic response(s) can be easily manipulated. Transgenic mice expressing the measles virus receptor will be useful for studying measles virus pathogenesis including virulence, tissue tropism, eliciting immune responses in the presence or absence of adoptively transferred antibodies, for mapping human HLA restricted CTL epitopes using double transgenic mice expressing CD46 and HLA class I molecules, creating a subunit vaccine and for testing of newly developed vaccines.

麻疹病毒在非洲造成严重的发病率和死亡率， 人口。 尽管减毒疫苗成功， 每年仍有100多万儿童死于这种病毒。问题 由于疫苗对在接种前感染的儿童无效， 他们九个月大的时候 据信，这是因为 母体抗体和/或病毒诱导的免疫抑制或其他 接种疫苗后的远期影响此外，麻疹会导致 超急性过敏性疾病和慢性持续性（亚急性） 硬化性全脑炎）神经元疾病。 然而我们的 对麻疹分子特征的认识和认识 病毒，免疫和免疫病理反应， 感染和疫苗接种以及病毒如何在神经元中持续存在， 不太了解。 为了解麻疹病毒感染的发病机制 并鉴定免疫保护反应，包括B和T 细胞表位，我们将开发和利用一种新的小动物， 模型我们和我们的同事已经确定了假定受体 麻疹病毒膜辅因子蛋白（MCP，CD 46）。 麻疹病毒对人类和猿类是允许的，但对鼠类是不允许的 细胞 然而，鼠MC 57和3 T3细胞几乎不允许 麻疹病毒变得宽容，并产生后代病毒时， 稳定表达四种CD 46同种型中的任何一种，尽管同种型 BC 1和BC 2与较高程度的合胞体 阵我们建议在小鼠中表达CD 46的cDNA， 自己的，β肌动蛋白（通用）和细胞特异性（神经元特异性 烯醇化酶，RIP）启动子。这将使我们能够生成一个 麻疹病毒的转基因模型在一个小的，便宜的， 基因可操纵的宿主，小鼠，其免疫 可以容易地操纵响应。 表达麻疹病毒受体的转基因小鼠将是 可用于研究麻疹病毒致病性包括毒力， 组织嗜性，在存在或 不存在过继转移抗体，用于人类HLA定位 限制性CTL表位，使用双转基因小鼠表达 CD 46和HLA I类分子，创造了一种亚单位疫苗， 测试新开发的疫苗。